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Confocal Microscopy Image Gallery
Madin-Darby Ovine Kidney Epithelial Cells (MDOK Line)
The MDOK cell line was established by S. H. Madin and N. B. Darby, Jr. from normal male ovine kidney tissue. Testing has demonstrated that the cells are susceptible to sheep bluetongue virus, infectious bovine rhinotracheitis, and the Indiana and New Jersey strains of vesicular stomatitis.
MDOK cells exhibit many characteristics typical of epithelial cells. In the body, epithelial cells are generally organized into sheet-like barriers that encase the organs and protect them from foreign substances. The cells often function in more than one way, however, and may be important for absorption and secretion as well as protection.
The sheep kidney, and the kidneys of other animals, is comprised of an outer cortex and an inner medulla. There is generally a large quantity of blood contained in the cortex, which renders the region of the kidney dark brownish-red in color. The abundance of renal corpuscles in the cortex also gives it a distinctive granular appearance. Due to the significantly reduced amount of blood in the medulla of the kidney, the inner section of the organ is much paler than the cortex. The medulla is organized into renal pyramids, which are striated sections of tissue separated form one another by the cortical columns. Located at the apex of each renal pyramid is a renal papilla, which features numerous small passages, known as the ducts of Bellini. The minor calyces that surround the apices come together in a series of major calyces, which empty urine into the renal pelvis, from which it is eventually excreted into the ureter and passed from the body.
The adherent monolayer culture of Madin-Darby ovine kidney cells illustrated above was immunofluorescently labeled with primary mouse anti-oxphos complex V inhibitor protein antibodies, followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 488. The culture was subsequently stained with Alexa Fluor 594 conjugated to phalloidin to reveal details of the filamentous actin network, and TO-PRO-3 (pseudocolored cyan) for DNA in the nucleus. Images were recorded with a 60x oil immersion objective using a zoom factor of 2.5 and sequential scanning with the 488-nanometer spectral line of an argon-ion laser, the 543-nanometer line from a green helium-neon laser, and the 633-nanometer line of a red helium-neon laser. During the processing stage, individual image channels were pseudocolored with RGB values corresponding to each of the fluorophore emission spectral profiles unless otherwise noted above.
Additional Confocal Images of Madin-Darby Ovine Kidney Epithelial (MDOK) Cells
Madin-Darby Ovine Kidney Cells Stained with Alexa Fluor Dyes and TO-PRO-3 - A culture of MDOK cells was labeled with TO-PRO-3 (pseudocolored blue), which exhibits affinity for DNA in the cell nucleus, and Alexa Fluor 594 conjugated to phalloidin, which binds to the cytoskeletal filamentous actin network. The epithelial cell culture was also probed immunofluorescently with primary anti-oxphos complex V inhibitor protein monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 488.
Visualizing Mitochondria in MDOK Cell Cultures with Anti-OxPhos Antibodies - The mitochondrial network in cultured Madin-Darby ovine kidney cells was immunofluorescently labeled with primary anti-oxphos complex V inhibitor protein monoclonal antibodies followed by goat anti-mouse Fab fragments conjugated to Alexa Fluor 488. In addition, the F-actin network was targeted with Alexa Fluor 594 (pseudocolored lavender) conjugated to phalloidin, and the nucleus was counterstained with TO-PRO-3.
Immunofluorescently Targeting the Golgi Complex and Nuclear Histone Proteins in Sheep Kidney Cells - The nucleus and Golgi complex in the MDOK epithelial cell culture shown in this confocal image were labeled in a double immunofluorescence experiment with mouse anti-histones (pan) and rabbit anti-giantin primary antibodies. The antibody targets were visualized with goat secondary antibodies conjugated to Texas Red and Alexa Fluor 488, respectively.
Distribution of F-Actin and Mitochondria in MDOK Cell Cultures - A log phase culture of Madin-Darby ovine kidney cells was labeled for F-actin and the intracellular mitochondrial network with Alexa Fluor 568 (pseudocolored blue) conjugated to phalloidin and MitoTracker Red CMXRos, respectively. The high-affinity DNA probe DRAQ5 (pseudocolored green) was utilized to counterstain cell nuclei.
Contributing Authors
Nathan S. Claxton, Shannon H. Neaves, and Michael W. Davidson - National High Magnetic Field Laboratory, 1800 East Paul Dirac Dr., The Florida State University, Tallahassee, Florida, 32310.